Intracellular labeling of single cortical astrocytes in vivo
Intracellular labeling of single cortical astrocytes in vivo
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DOI：
10.1016/j.jneumeth.2007.06.021
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发表时间：
2007-10-15
影响因子：
3
通讯作者：
Hirase, Hajime
中科院分区：
文献类型：
--
作者：
Mishima, Tsuneko;Sakatani, Seiichi;Hirase, Hajime
传统上，神经胶质细胞被认为在中枢神经系统中发挥支持作用。最近的实验证据表明，神经胶质细胞参与了神经信息处理，因此有必要在成熟的大脑中监测体内胶质细胞的生理学。同时，由于至少存在几种不同的神经胶质细胞，因此对记录的神经胶质细胞进行鉴定和分类是至关重要的。过去的研究已经实现了使用锐利玻璃微电极对胶质细胞进行在体细胞内电生理记录，但由于技术上的困难，记录的细胞的形态恢复和鉴定还很难进行。我们证明，使用大片段生物素化葡聚糖胺(BDA)是一种有效的方法来标记单个胶质细胞与尖锐的微电极在体。此外，示踪信号的放大是通过亲和素生物素化的辣根过氧化物酶大分子复合体(ABC)和基于酪胺的方法相结合实现的，从而使多重免疫组织化学成为可能。使用本研究中描述的方法，我们已经成功地记录并标记了包括星形胶质细胞、少突胶质细胞和小胶质细胞在内的大脑皮质细胞。(C)2007 Elsevier B.V.保留所有权利。
Glial cells have traditionally been considered to play supportive roles in the central nervous system. As recent experimental evidence suggests glial cells' participation in neural information processing, there has been a need to monitor the physiology of glial cells in vivo in the matured brain. Concurrently, identification and classification of the recorded glial cells is essential as there are at least several different kinds of glial cells. Past studies have achieved in vivo intracellular electrophy siological recording of glial cells using sharp glass microelectrodes, however, morphological recovery and identification of the recorded cells have hardly been done, due to technical difficulties. We demonstrate that use of large fragment biotinylated dextran amine (BDA) is an effective way to label a single glial cell recorded with a sharp microelectrode in vivo. Furthermore, the tracer signal amplification was achieved by a combination of avidin biotinylated horseradish peroxidase macromolecular complex (ABC) and tyramide-based methods, making multiple immunohistochemistry feasible. Using the method described in this study, we have successfully recorded and labeled cortical galial cells including astrocytes, oligodendrocytes, and microglia. (C) 2007 Elsevier B.V. All rights reserved.